E2F Reporter Lentivirus
Fluorescent E2F Reporter Lentivirus: High-quality, fluorescent lentiviral transcription factor (TF) reporter system that provides sensitive fluorescent readout for human/mouse E2F transcription factors (E2F) activity in transduced cells. The E2F reporter is constructed with tandem repeats of consensus E2F DNA-binding elements to read out the activities of E2F transcription factors in mammalian cells. As a member of the E2F family of transcription factors, E2F2 often act as transcriptional activator to activate certain target genes such as cyclin A and E in the cell cycle entry for proliferation. Tumor suppressor Retinoblastoma protein (Rb) in unphosphorylated form binds to E2F2 in G0/G1, forming a complex that actively represses E2F2-responsive genes. CDKs phosphorylate Rb proteins causing the release and accumulation of sequestered E2F2. E2F2 overexpression promotes the development and progression of tumors such as breast, lung cancer, and liver cancer. The E2F reporter lentivirus is purified by PEG precipitation and sucrose gradient centrifugation, and is ideal for studying E2F transcription factor activity in difficult-to-transfect cells including primary and/or thawed cells.
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Ultra-high concentration virus can be provided upon request in your choice of medium and volume for in vivo injection. We can provide custom reporter genes (such as SEAP, YFP, secreted Gaussia Luc, Renilla Luc, etc.) and selection markers (hygromycin, bleomycin, etc.) upon request.
- High Sensitivity – LipExoGen TF Reporter lentiviral particles are made using a novel vector platform based on the third generation system. The transcription factor’s response elements are arranged as DNA tandem repeats upstream of the minimal TATA promoter-driven reporter, and downstream of an optimized minimal enhancer (pc) of the human CMV promoter. When the signal pathway/TF is activated, the mini enhancer synergizes with TF binding to the response elements (up to 8 repeats in some products, depending on strength of reporter activation) to amplify expression of the fluorescent (GFP/RFP) or luciferase (Luc) reporter, with minimal enhancement of background. As a result, the reporter system provides a highly sensitive readout for signaling pathway or specific transcription factor activation in human and mouse cells.
- Easily Establish Stable Reporter Cell Lines – The reporter lentiviral particles are ultra-purified and concentrated to high-titer by PEG purification and sucrose gradient centrifugation to allow for efficient transduction of difficult-to-transfect cells, including primary and/or freeze-thawed cells. Stable cell lines are easily generated through puromycin or blasticidin selection.
- Discovery Made Easy – Signal pathway or specific transcription factor activity can be detected by fluorescence, making LipExoGen TF Reporter lentiviral particles more practical than traditional luciferase reporters and/or biochemical assays. Pathway/TF activation can be readout directly by fluorescence microscopy in living cell cultures, thus paving the way for unexpected discoveries.
- Readout On Flow – Fluorescent reporter activation can also be readout by flow cytometry, providing more versatility in data acquisition for labs with different instruments.
- Best Value – LipExoGen lentiviral particle products are made using optimized lentiviral vectors developed in-house, which allows us to provide the highest quality products while retaining competitive prices. These high-titer lentiviral particles feature a highly sensitive fluorescent reporter system and have been validated to read out the indicated transcription factor activity.
- Same Cost For Custom Lentivirus – You can easily request any combination of reporter (GFP/RFP/Luc) and selection marker (puromycin/blasticidin) for this product, without additional cost, by contacting us. To view our complete list of vectors, click here.
Figure 1 (thumbnail). HEK293FT cells were co-transfected with E2F-TAG-Puro plus either control plasmid (Vector, left) or expression plasmid for human E2F2 (E2F2, right). Fluorescence microscopy images were taken 24 hrs later to assess GFP reporter expression in the live cells.
Figure 2. HEK293FT cells were transfected with E2F-TAG-Puro in combination with an empty vector (pcDNA) or cDNA for full length wild type human E2F2 or an an E2F2 TAD mutant for 24 hours before fluorescence microscopy.
Have questions about this product? Send us a form and we’ll reply the same day: Contact Us
|E2F transcription factors|
|E2F consensus DNA-binding elements|
|Monitor E2F transcription factor activity in mammalian cells, screen for activators or inhibitors of E2F pathway. The fluorescent reporter enables convenient readout using flow cytometry, fluorescence microscopy, etc.|
Renilla Luciferase Internal Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven Renilla luciferase (RLuc).
Reporter Negative Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven reporter. The construct is the same as the TF Reporters except that it lacks the transcriptional response elements which drive signal pathway/TF-specific reporter expression. The reporter negative control lentiviral particles allow to establish a baseline for background reporter activity and determine specificity of any treatments to activate the reporter.
Reporter Positive Control Lentivirus – Ready-to-transduce lentiviral particles with constitutive expression of the reporter. The reporter positive control lentivirus is useful for transduction optimization studies, especially if the cells are very sensitive or difficult-to-transduce.
If you require a modification to one of our products, such as a change in reporter or other vector component, contact us. We provide fast and efficient services for custom cloning and lentiviral particles on demand, usually for the same or comparable price as the listed item.
Or, send us your cells and we will establish a stable reporter cell line for you using this product. Learn more.
|E2F transcription factor 1|
|Homo sapiens/mus musculus|
|Alias||RBP3; E2F-1; RBAP1; RBBP3|
|Gene IDs||HGNC:HGNC:3113 Ensembl:ENSG00000101412 MIM:189971|
|Entrez Gene Summary||“The protein encoded by this gene is a member of the E2F family of transcription factors. The E2F family plays a crucial role in the control of cell cycle and action of tumor suppressor proteins and is also a target of the transforming proteins of small DNA tumor viruses. The E2F proteins contain several evolutionally conserved domains found in most members of the family. These domains include a DNA binding domain, a dimerization domain which determines interaction with the differentiation regulated transcription factor proteins (DP), a transactivation domain enriched in acidic amino acids, and a tumor suppressor protein association domain which is embedded within the transactivation domain. This protein and another 2 members, E2F2 and E2F3, have an additional cyclin binding domain. This protein binds preferentially to retinoblastoma protein pRB in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent/independent apoptosis. [provided by RefSeq, Jul 2008]”|